Application of BGT226 maleate in the preparation of anti-novel coronavirus drugs

Abstract

This invention discloses the application of BGT226 maleate in the preparation of drugs against the novel coronavirus, belonging to the field of antiviral drug technology. The application of BGT226 maleate in the preparation of drugs against the novel coronavirus shows that BGT226 maleate has an IC50 of 0.116 μM against the original SARS-CoV-2 strain and an IC50 of 0.084 μM against the Omicron variant (JN.1), and exhibits good biocompatibility. This reveals its potential for developing drugs against novel coronavirus infection, providing a new direction and possibility for the treatment of the novel coronavirus and its variants.

Description

Technical Field

[0001] This invention relates to the field of antiviral drugs, and in particular to the application of BGT226 maleate in the preparation of drugs against the novel coronavirus. Background Technology

[0002] Current strategies for developing drugs against the novel coronavirus mainly include targeted drug design and development against key viral molecules (such as Mpro, RdRP, PLpro, etc.) and host factors (such as ACE2, TMPRSS2, etc.), and drug screening based on viral infection models. The former involves developing inhibitors such as small molecules, peptides, and aptamers through virtual or experimental screening based on molecular interactions, and developing neutralizing antibodies through antigen-immune screening or antibody gene sequencing. The latter involves screening for drugs or compounds that can inhibit viral replication by testing various existing drug and compound libraries (including libraries of known active compounds and natural product compounds).

[0003] BGT226 maleate is the maleate-modified form of BGT226, which improves the water solubility of the drug molecule. BGT226 is a PI3K / mTOR dual inhibitor developed by Novartis (IC50 values ​​for PI3Kα, PI3Kβ, and PI3Kγ are 4 nM, 63 nM, and 38 nM, respectively). Since the PI3K / Akt / mTOR pathway plays a crucial role in cell proliferation and survival, inhibiting this pathway induces cell cycle arrest and apoptosis, thus producing a cytotoxic effect on tumor cells. Studies have confirmed that BGT226 has strong growth-inhibiting activity against various tumor cell types. Phase I clinical trials showed good pharmacokinetics and a suitable dose range. However, the therapeutic effect of BGT226 maleate in novel coronavirus infection has not yet been investigated. Summary of the Invention

[0004] The purpose of this invention is to provide the application of BGT226 maleate in the preparation of anti-novel coronavirus drugs, which is a new application of BGT226 maleate in novel coronavirus infection and has good development prospects.

[0005] To achieve the above objectives, this invention provides the application of BGT226 maleate in the preparation of drugs against the novel coronavirus. The molecular formula of BGT226 maleate is C2. 32 H 29 F3N6O6, the structural formula is shown below:

[0006] .

[0007] Preferably, the novel coronavirus includes the original SARS-CoV-2 strain and the Omicron variant JN.1.

[0008] Preferably, the IC50 of BGT226 maleate against the original SARS-CoV-2 strain is 0.116 μM.

[0009] Preferably, the IC50 of BGT226 maleate against the Omicron mutant (JN.1) is 0.084 μM.

[0010] Therefore, the application of BGT226 maleate in the preparation of drugs against the novel coronavirus in this invention has shown that BGT226 maleate has an IC50 of 0.116 μM against the original SARS-CoV-2 strain and an IC50 of 0.084 μM against the Omicron variant (JN.1), and exhibits good biocompatibility. This reveals that it can be used to develop drugs against novel coronavirus infection and is a potential drug for treating novel coronavirus infection, providing a new direction and possibility for the treatment of novel coronavirus and its variants.

[0011] The technical solution of the present invention will be further described in detail below with reference to the accompanying drawings and embodiments. Attached Figure Description

[0012] To more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings used in the description of the embodiments of the present invention will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0013] Figure 1 A statistical chart showing the percentage of virus inhibition in different treatment groups;

[0014] Figure 2 Morphological observations of cells in different treatment groups. Detailed Implementation

[0015] The technical solution of the present invention will be further described below with reference to the accompanying drawings and embodiments.

[0016] To make the objectives, technical solutions, and advantages of this application clearer, more thorough, and more complete, the technical solutions of the present invention will be clearly and completely described below with reference to the accompanying drawings and embodiments. The following detailed descriptions are all illustrations of embodiments, intended to provide further detailed explanation of the present invention. Unless otherwise specified, all technical terms used in this invention have the same meaning as commonly understood by one of ordinary skill in the art to which this application pertains.

[0017] The instruments, equipment, reagents, and materials used in the examples were all obtained commercially.

[0018] The molecular formula of the BGT226 maleate used is: C 32 H 29 F3N6O6, structural formula is:

[0019] .

[0020] Example

[0021] Constructing a drug screening model against the novel coronavirus:

[0022] (1) African green monkey kidney Vero cells were seeded in DMEM medium and cultured in a 37°C, 5% CO2 incubator. One day before drug screening, 2.5 × 10⁻⁶ cells were seeded... 4 A sufficient number of Vero cells were seeded into 96-well plates, and their density was increased to over 90% by the second day. The initial concentration of the small molecule kinase inhibitor library was 10 mM, which was first diluted to 100 μM with DMEM medium, and then serially diluted 2-fold to seven other concentrations. 8 μL of the diluted drug was pipetted into each well of the Vero cell culture plate, with a medium concentration of 150 μL per well.

[0023] (2) In a biosafety level 3 laboratory, novel coronavirus infection was performed on the original SARS-CoV-2 strain and the Omicron variant (JN.1). The original virus solution was diluted with DMEM medium at an appropriate ratio. 50 μL of the diluted virus solution was added to the culture wells to achieve a final infection titer of 100 times TCID50. The final drug concentrations were 4, 2, 1, 0.5, 0.25, 0.125, 0.0625 and 0.03125 μM, respectively. A negative control without virus infection and a positive control with virus but without drug were set up, with 3 replicates for each.

[0024] (3) Three days after viral infection, the viral load in the culture supernatant was detected using a novel coronavirus RNA detection kit (Shanghai Berger Medical Technology Co., Ltd., catalog number: ZC-HX-201-02, National Medical Device Registration Certificate No. 20203400065) to evaluate the inhibitory effect of the drug on viral replication. At the same time, the cytopathic effect was observed under a microscope. If high concentrations of the drug caused cell death, the drug concentration was considered ineffective.

[0025] The testing process is as follows:

[0026] After culture, 100 µL of culture supernatant was collected and viral nucleic acid was extracted using a magnetic bead nucleic acid extraction kit and an automated nucleic acid extractor (Shanghai ZJ Biotechnology). The viral RNA level was detected using a novel coronavirus nucleic acid detection kit (fluorescent PCR method): Specific primers and probes were designed for the ORF1a / b and N genes of the novel coronavirus RNA (NC_045512.2) using a one-step RT-PCR method combined with TaqMan technology. The ORF1a / b gene probe was labeled with FAM fluorescein, the N gene probe with VIC fluorescein, and the internal control probe with ROX fluorescein. After 45 cycles of PCR denaturation (97℃, 5s), annealing, and extension (58℃, 30s), the light signals emitted after hydrolysis of the probes labeled with different fluoresceins were collected by the PCR instrument, displaying amplification curves on the instrument, thus enabling the detection of the novel coronavirus ORF1a / b and N genes.

[0027] The level of the virus is represented by the Ct value, through 2 -ΔΔCt The method calculates the relative amount of viral nucleic acid in each drug-treated group and the drug-free group to obtain the percentage of viral inhibition, and evaluates the drug's activity against the novel coronavirus.

[0028] Virus inhibition percentage such as Figure 1 As shown, the microscopic images of the cells are as follows: Figure 2 As shown, 2, 1, 0.5, 0.25, 0.125, 0.0625, and 0.03125 μM BGT226 maleate can inhibit viral replication without significantly altering cell morphology. Although 4 μM BGT226 maleate inhibited viral replication, it significantly altered cell morphology, indicating that this drug concentration caused considerable cytotoxicity and therefore was not included in the effective concentration.

[0029] In summary, the IC50 of BGT226 maleate against the original SARS-CoV-2 strain is 0.116 μM, and the IC50 against the Omicron variant (JN.1) is 0.084 μM.

[0030] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention and not to limit them. Although the present invention has been described in detail with reference to preferred embodiments, those skilled in the art should understand that modifications or equivalent substitutions can still be made to the technical solutions of the present invention, and these modifications or equivalent substitutions cannot cause the modified technical solutions to deviate from the spirit and scope of the technical solutions of the present invention.

Claims

1. Use of BGT226 maleate in the preparation of a drug for resisting the novel coronavirus, characterized in that, The molecular formula of BGT226 maleate is C 32 H 29 F3N6O6, and the structural formula is as follows: ； The novel coronavirus is Omicron variant JN.1 of SARS-CoV-2 original strain. 2.The use of the BGT226 maleate salt according to claim 1 in the preparation of an anti-2019-nCoV drug, characterized in that: The IC50 of BGT226 maleate against Omicron variant JN.1 is 0.084 µM.

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